Institute of Orthopedic Research, General Hospital of the People's Liberation Army, Beijing 100853, China.
J Exp Clin Cancer Res. 2011 Feb 26;30(1):24. doi: 10.1186/1756-9966-30-24.
The immune factors heat shock protein (HSP)/peptides (HSP/Ps) can induce both adaptive and innate immune responses. Treatment with HSP/Ps in cancer cell-bearing mice and cancer patients revealed antitumor immune activity. We aimed to develop immunotherapy strategies by vaccination with a mixture of HSP/Ps (mHSP/Ps, HSP60, HSP70, Gp96 and HSP110) enhanced with cyclophosphamide (CY) and interleukin-12 (IL-12).
We extracted mHSP/Ps from the mouse sarcoma cell line S180 using chromatography. The identity of proteins in this mHSP/Ps was assayed using SDS-PAGE and Western blot analysis with antibodies specific to various HSPs. BALB/C mice bearing S180 cells were vaccinated with mHSP/Ps ×3, then were injected intraperitoneally with low-dose CY and subcutaneously with IL-12, 100 μg/day, ×5. After vaccination, T lymphocytes in the peripheral blood were analyzed using FACScan and Cytotoxicity (CTL) was analyzed using lactate dehydrogenase assay. ELISPOT assay was used to evaluate interferon γ (IFN-γ), and immune cell infiltration in tumors was examined in the sections of tumor specimen.
In mice vaccinated with enhanced vaccine (mHSP/Ps and CY plus IL-12), 80% showed tumor regression and long-term survival, and tumor growth inhibition rate was 82.3% (30 days), all controls died within 40 days. After vaccination, lymphocytes and polymorphonuclear leukocytes infiltrated into the tumors of treated animals, but no leukocytes infiltrated into the tumors of control mice. The proportions of natural killer cells, CD8+, and interferon-γ-secreting cells were all increased in the immune group, and tumor-specific cytotoxic T lymphocyte activity was increased.
In this mice tumor model, vaccination with mHSP/Ps combined with low-dose CY plus IL-12 induced an immunologic response and a marked antitumor response to autologous tumors. The regimen may be a promising therapeutic agent against tumors.
免疫因素热休克蛋白（HSP）/肽（HSP/Ps）可诱导适应性和固有免疫反应。在荷瘤小鼠和癌症患者中用 HSP/Ps 治疗显示出抗肿瘤免疫活性。我们旨在通过用环磷酰胺（CY）和白细胞介素-12（IL-12）增强的 HSP/Ps（mHSP/Ps、HSP60、HSP70、Gp96 和 HSP110）混合物进行疫苗接种来开发免疫治疗策略。
我们使用色谱法从鼠肉瘤细胞系 S180 中提取 mHSP/Ps。使用 SDS-PAGE 和针对各种 HSP 的特异性抗体进行 Western blot 分析来检测该 mHSP/Ps 中的蛋白质的身份。荷 S180 细胞的 BALB/C 小鼠接种 mHSP/Ps×3 次，然后腹腔内注射低剂量 CY，皮下注射 IL-12，每天 100μg，共 5 天。接种后，使用 FACScan 分析外周血中的 T 淋巴细胞，并用乳酸脱氢酶测定法分析细胞毒性（CTL）。使用 ELISPOT 测定法评估干扰素 γ（IFN-γ），并检查肿瘤标本切片中的免疫细胞浸润。
在接种增强疫苗（mHSP/Ps 和 CY 加 IL-12）的小鼠中，80%显示肿瘤消退和长期存活，肿瘤生长抑制率为 82.3%（30 天），所有对照组均在 40 天内死亡。接种后，淋巴细胞和多形核白细胞浸润到治疗动物的肿瘤中，但对照组的肿瘤中没有白细胞浸润。免疫组的自然杀伤细胞、CD8+和干扰素-γ分泌细胞的比例均增加，并且肿瘤特异性细胞毒性 T 淋巴细胞活性增加。
在该小鼠肿瘤模型中，用 mHSP/Ps 联合低剂量 CY 加 IL-12 接种可诱导免疫反应，并对自体肿瘤产生显著的抗肿瘤反应。该方案可能是一种有前途的肿瘤治疗剂。
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